Overview

  • Product nameMitoTox™ Complex V OXPHOS Activity Assay Kit
    See all Complex V kits
  • Detection methodColorimetric
  • Sample type
    Inhibitor compounds
  • Assay typeDirect
  • Assay time
    4h 00m
  • Species reactivity
    Reacts with: Rat, Cow, Human
    Does not react with: Mouse
  • Product overview

    MitoTox™ Complex V OXPHOS Activity Assay Kit (ab109907) is designed for testing the direct inhibitory effect of compounds on Complex V activity in only 4 hours. Complex V extracted from the provided bovine heart mitochondria (a rich source of Complex V) is immunocaptured by specific antibodies on the plate. Complex V activity can be observed as decrease in absorbance at OD 340 nm. The intra-assay and inter-assay variation of this assay are both < 10%.


    Inhibitory effects of compounds on Complex I activity can be tested in two different ways: 1. Screening format, where up to 23 compounds can be tested at a single concentration in triplicate; 2. Dose response (IC50) format, where two compounds known to affect Complex V activity can be tested at 11 different data points in triplicate.


    Testing for mitochondrial function has become a key aspect of drug discovery. Mitochondria can be affected by drug treatment, resulting into cardio- and hepatotoxic side effects that can lead to drug withdrawal from the market. Therefore, there is increasing emphasis on testing the impact on mitochondria early on in the drug development process to reduce failure rates during preclinical and clinical phases. 

  • Notes

    Store Phospholipids, Bovine heart mitochondria and Activity Buffer at -80°C. Store all other components at 4°C.

  • Tested applicationsSuitable for: Functional Studiesmore details
  • PlatformMicroplate

Properties

    Applications

    Our Abpromise guarantee covers the use of ab109907 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    Application Abreviews Notes
    Functional Studies Use at an assay dependent dilution.

    MitoTox™ Complex V OXPHOS Activity Assay Kit images

    • Dose response curve for aurovertin, a non-competitive inhibitor of Complex V.

    • Typical dose response curve for oligomycin. Assay was performed following the Dose Response Assay Procedure using oligomycin, a well known Complex V inhibitor. Oligomycin was prepared in DMSO to generate a 10 mM stock. Starting with a 50 µM final concentration in well, 1:10 serial dilutions of oligomycin were generated.

    Protocols

    References for MitoTox™ Complex V OXPHOS Activity Assay Kit (ab109907)

    This product has been referenced in:
    • Xu Z  et al. Acute upregulation of neuronal mitochondrial type-1 cannabinoid receptor and it's role in metabolic defects and neuronal apoptosis after TBI. Mol Brain 9:75 (2016). Read more (PubMed: 27485212) »
    • Quan X  et al. Essential role of mitochondrial Ca2+ uniporter in the generation of mitochondrial pH gradient and metabolism-secretion coupling in insulin-releasing cells. J Biol Chem 290:4086-96 (2015). Read more (PubMed: 25548283) »

    See all 6 Publications for this product

    Product Wall

    Thank you for contacting us.

    Unfortunately none of our products measure the forward reaction. As far as I know there are no companies selling kits measuring the forward reaction of the ATP synthase I am afraid.

    I am sorry I could no...

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    The kits that are the best suited for your experiment are listed below. I am having our datasheets team update the species reactivity.
    ab109903, Complex I; reactivity = Human, mouse, rat, bovine
    http://www.abcam.com/mitotox%E2%84%A2-complex-i...

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    Thank you for contacting us.

    The bovine mitochondria is in Tris, sucrose, EDTA buffer pH 7.8


    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
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    Product code: 109714 Inquiry: Hi Abcam, Do you know if this kit will immunocapture the ATP synthase enzyme from C. elegans? Unlikely unfortunately – this kit was tested positive with human, rat and cow. Mouse was negative. I think it is unl...

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    Unfortunately not. It comes with a sample (bovine) which cannot be substituted for mouse.

    The rate quoted in the booklet is mOD/min. I think this customer is quoting OD. Therefore 0.12/18 is 0.0067 OD/min = 6.7 mOD/min

    Changing the window analyzed to identify the steepest straight line slope is necessary and then to compare all sa...

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    Thank you for your enquiry.

    The negative control chosen for ab109907 MitoTox™ Complex V OXPHOS Activity Microplate Assaywould depend on whatyou want to accomplish and obtain from the experiment.

    Some people test the specific...

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"